Integrated biocontainment cell sorter

ABSTRACT

Disclosed is an integrated biocontainment cell sorter that isolates portions of the cell sorter that can create contamination. Two containment systems are utilized. A main cabinet containment system contains input samples. An aerosol management containment area includes a nozzle chamber with a nozzle and a sort chamber with sort plates and collection media that collect a droplet stream from the nozzle. The main cabinet is maintained at a first low pressure and clean air is recirculated under a positive pressure. The aerosol management containment area is kept at a second low pressure, which is lower than the first pressure, so that contamination does not leak from the aerosol management containment area into the main cabinet containment area. A sliding sash window is located over an access opening in the main cabinet and can be moved to access different portions of the main cabinet without changing the substantially constant first low pressure in the main cabinet.

CROSS-REERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/910,538, “Integrated Biocontainment Cell Sorter” (filed Jun. 24,2020) (now allowed), which claims benefit to U.S. patent applicationSer. No. 62/866,759, “Integrated Biocontainment Cell Sorter” (filed Jun.26, 2019), the contents of which foregoing applications are incorporatedherein by reference in their entireties for any and all purposes.

BACKGROUND

Cell sorter flow cytometers have become an important laboratory tool.Cell sorters are capable of identifying certain types of biologicalcells and separating those cells from other cells. Commercial uses ofcell sorters have also been implemented in several industries. There aremany other uses of cell sorters, such as identifying and isolatingvarious types of cells for laboratory applications. As such, cellsorters have many different and varied uses and applications.

SUMMARY

An embodiment of the present invention my therefore comprise anintegrated biocontainment cell sorter flow cytometer comprising: a maincabinet of the integrated biocontainment cell sorter that is nothermetically sealed; an input sample area disposed in the main cabinet;a moveable partition in the main cabinet that moves in an access openingof the main cabinet, the moveable partition covering a constant amountof area of the access opening as the moveable partition is moved in theaccess opening which leaves a constant amount of area of the accessopening that is not covered by the moveable partition and is open, asthe moveable partition is moved in the access opening; a first fan thatdraws air from the main cabinet to create a first low pressure in themain cabinet that is substantially constant as the moveable partition ismoved in the access opening, which limits contaminated air in the maincabinet from escaping from the main cabinet; an aerosol managementcontainment area that is not hermetically sealed, which is disposed inthe main cabinet, the aerosol management containment area havingopenings that are connected to the main cabinet so that the aerosolmanagement containment area is disposed in and subject to the first lowpressure, the aerosol management containment area having a nozzle thatcreates a droplet stream containing sample cells, sort plates thatseparate the droplet stream into deflected stream and collection media,that collect the deflected streams; a second fan that draws air from theaerosol management containment area to create a second low pressure inthe aerosol management containment area that is lower than the first lowpressure which causes air from the main cabinet to flow from the maincabinet to the aerosol management containment area and limitscontaminated air from flowing from the aerosol management containmentarea into the main cabinet; optical excitation devices located outsideof the main cabinet and the aerosol management containment area to allowaccess to the optical excitement devices without accessing the maincabinet or the aerosol management containment area.

Another embodiment of the present invention may therefore comprise amethod of containing cells in an integrated biocontainment cell sortercomprising: providing a main cabinet containment area that contains aninput area for sample cells to be sorted; generating a first lowpressure in the main cabinet using a first fan that draws air from themain cabinet and air from outside the cabinet; generating a second lowpressure in an aerosol management containment area, disposed in the maincabinet, using a second fan that draws air from the main cabinet and theaerosol management containment area through openings in the aerosolmanagement containment area to create the second low pressure in theaerosol management containment area that is lower than the first lowpressure; enclosing input cell samples in the main cabinet that is nothermetically sealed; enclosing a nozzle, sort plates, collection mediaand any droplet stream, created by the nozzle, in an aerosol managementcontainment area that is not hermetically sealed; locating opticalexcitation devices outside of the main cabinet and the aerosolmanagement containment area for easy access for adjustment andmaintenance of the optical excitation devices.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic side cutaway view illustrating various parts ofone embodiment of an integrated biocontainment cell sorter.

FIG. 2 is a schematic illustration of the Aerosol Management System(AMS) of FIG. 1 , including the nozzle chamber and sort chamber andvarious other parts of an embodiment of an integrated biocontainmentcell sorter.

FIG. 3 is a more detailed side cutaway view illustrating the nozzlechamber and the sort chamber of FIG. 2 .

FIG. 4 is a detailed front view of an embodiment of an integratedbiocontainment cell sorter showing the nozzle chamber and sort chamberof FIGS. 2 and 3 .

FIG. 5 is a perspective view of an implementation of the integratedbiocontainment cell sorter that is illustrated in FIGS. 1-4 .

DETAILED DESCRIPTION OF THE EMBODIMENTS

FIG. 1 is a schematic diagram of one embodiment of an integratedbiocontainment system 100. The system is comprised of a main cabinetcontainment system 101 and aerosol management system chambers 140, theAMS intake duct 138, AMS HEPA filter 141, AMS fan 144 and AMS exhaustduct 147, which are collectively referred to as the aerosol managementsystem (AMS) 149. The aerosol management system (AMAS) 149 is containedin or connected to the main cabinet containment system 101. Neither themain cabinet containment system 101 nor the aerosol management system(AMS) 149 are sealed systems. Rather, they rely on movement of air byfan 122, in the main cabinet containment system 101, and AMS fan 144 inthe aerosol management system (AMS) 149, to create low pressure, so thatharmful and toxic materials are not spread outside of each of thesecontainment systems. For example, a first low pressure is created in thework area 104 of the main cabinet containment system 101 as a result offan 122 extracting air from both the work area 104 and air from outsideof the main cabinet containment system 101, which is shown by airstreams 116. In this manner, a first low pressure is created in the workarea 104 where potentially toxic or dangerous materials may exist. Aslong as the fan 122 is able to pull sufficient air through the work area104, dangerous materials will not escape from the work area 104 to anarea outside the main cabinet containment system 101. Dangerousmaterials primarily exist as aerosols that contain sample cells, Samplecells are mixed with sheath fluid and passed through a nozzle 146.Normally, the nozzle creates a nozzle stream 153 (FIG. 2 ) that breaksup into a droplet stream 155 (FIG. 2 ). If the nozzle becomes clogged,an aerosol can be created that contains sample cells. Also, if thenozzle stream 153 or the droplet stream 155 hit a hard surface, aerosolscan be created. The aerosols contain sample cells that should not beinhaled or ingested. For example, the sample cells may be cancer cells.

As also shown in FIG. 1 , air drawn by the fan 122 first passes throughthe HEPA filter 120 to remove any dangerous materials such as samplecells. Consequently, the air drawn by the fan 122 is clean air and thefan is not contaminated. The fan 122 forces the clean air through therecirculation duct 108 under positive pressure, which also remainsclean. The recirculation duct 108 and recirculation plenum do not haveto be sealed, even though they are under positive pressure, since theycontain clean air. Part of the air from the fan 122 is exhausted out ofthe exhaust vent 110, as illustrated by exhaust air 126. At the sametime, some of the air from the fan 122 is recirculated, as illustratedby recirculation air 124. Hence, the recirculation duct 108 circulatesclean air under positive pressure, and clean air is recirculated intothe recirculation plenum 102 under positive pressure, while theremaining air is exhausted out of the exhaust vent 110. The recirculatedair in the recirculation duct 108 and the recirculation plenum 102 isunder positive pressure and as such may leak from these ducts to theoutside air or to other parts of the integrated biocontainment system100. Since the air under positive pressure is clean air, there are noproblems with contamination, unlike many other containment systems. Therecirculated air in the recirculation plenum 102 passes through anairflow straightener 128. The airflow straightener 128 is a device thathas openings that cause the recirculated air to flow into the work area104 as a substantially uniform, laminar air flow 130 with lowturbulence. Low turbulence allows for maintenance of a uniform downwardvolume of air that prevents both contaminates from the inside of thecabinet from escaping the user access opening and prevents contaminatesfrom the outside of the cabinet from depositing on the products presentinside the cabinet.

The aerosol management system chambers 140 have openings that connect tothe main cabinet containment system 101, as also illustrated in FIG. 1 .The aerosol management system (AMS) 149 has a separate AMS HEPA filter141 and a separate AMS fan 144. AMS fan 144 draws air from the AMSintake duct 138, which is connected to the sort chamber 131. Since theaerosol management system chambers 140 are connected by openings to themain cabinet containment system 101, the aerosol management systemchambers 140 are already at the first low pressure that is maintained inthe main cabinet containment system 101. AMS fan 144 further lowers thepressure in the aerosol management system (AMS) 149 from the first lowpressure of the work area 104 to a second low pressure that is lowerthan the first low pressure. When the nozzle chamber door 136 is opened,or the sort chamber door 132 is opened, the second low pressure in theaerosol management system chambers 140 will equalize with the first lowpressure of the main cabinet containment system 101, So, as thepressures of the main cabinet containment system 101 and the aerosolmanagement system chambers 140 equalize as a result of either the nozzlechamber door 136 or the sort chamber door 132 being opened, airinitially flows from the main cabinet containment system 101 to theaerosol management system chambers 140 which prevents aerosols fromescaping the aerosol management system chambers 140. However, once thepressures are equalized, there can be a migration of aerosols from theaerosol management system chambers 140 to the main cabinet containmentsystem 101. Consequently, prior to opening either the nozzle chamberdoor 136 or the sort chamber door 132, the nozzle 146 is shut down andthe AMS fan 144 is operated at an increased speed for a time to evacuateall aerosols from the aerosol management system chambers 140. Duringoperation, with the nozzle chamber door 136 and the sort chamber door132 closed, the second low pressure air in the aerosol management systemchambers 140 draws air from the work area 104 of the main cabinetcontainment system 101 at AMS inlet 135 and AMS inlet 137 and AMS inlet160. In other words, air from the man cabinet containment system 101 inthe work area 104, which is at a first low pressure, is drawn into theaerosol management system chambers 140 since the second low pressure inthe aerosol management system chambers 140 is lower than the first lowpressure in the work area 104 while the nozzle chamber door 136 and sortchamber door 132 are closed. Again, this is a result of the AMS fan 144drawing air from the aerosol management system chambers 140 through theAMS HEPA filter 141 via AMS intake duct 138 to the outside environment.Clean air that has been filtered by the AMS HEPA filter 141 and drawn bythe AMS fan 144 is exhausted through the AMS exhaust duct 147 to theexhaust vent 110. Accordingly, the aerosol management system (AMS) 149is not only a parallel containment system to the main cabinetcontainment system 101, the aerosol management system chambers 140 areconnected by air inlets to the main cabinet containment system 101 tocreate a second lower pressure, which makes it doubly hard for dangerousmaterials located in the aerosol management system chambers 140 and thework area 104 to escape from the integrated biocontainment system 100.

As further illustrated in FIG. 1 , the aerosol management systemchambers 140 are carefully constructed to enclose portions of the cellsorter that produce hazardous particles and not enclose portions of thecell sorter that do not produce hazardous particles, to minimize thesize of the containment area and consequently minimize the size of theintegrated biocontainment system 100 for cell sorters. As shown in FIG.1 , the nozzle chamber 134 contains the nozzle 146 which is fed samplefluid 143 and sheath fluid 145. The nozzle 146 creates a nozzle stream153 (FIG. 2 ) from the sample fluid 143 and sheath fluid 145 that passesthrough the interrogation point 148 and through an opening 152 in theoptics mounting plate 150. Since the sample 143 may contain dangerousmaterials, e.g., dangerous cells that can be dispersed in aerosol formwhen nozzle 146 is clogged, the nozzle 146 is contained within thenozzle chamber 134 to prevent any dangerous aerosols from escaping theaerosol management system chambers 140. The optics mounting plate 150separates the nozzle chamber 134 from the sort chamber 131. Opening 152allows the interrogated droplet stream 155 (FIG. 2 ) to pass through theoptics mounting plate 150 to the sort plates 154. Each droplet of thedroplet stream 155 (FIG. 2 ) that is interrogated at interrogation point148 is then separated by the sort plates 154. The deflected dropletstreams 156 are then collected by the collection media 158. This isexplained in more detail in U.S. Pat. No. 8,557,587 issued on Oct. 15,2013 to Fox et al., which is specifically incorporated herein, byreference, for all that it discloses and teaches.

The main cabinet containment system 101 is primarily used to contain thesample input area 107 (FIG. 5 ) from the ambient air outside of the maincabinet containment system 101. The sample input area 107 is located inthe lower portion of the main cabinet containment system 101. Thesamples are placed in the sample input area 107 that is part of the workarea 104. The samples may comprise biohazardous material. They are firstprepared in a protected area such as a large dedicated biosafety cabinetthat is independent of the biocontainment cell sorter system disclosedherein. The cell samples are suspended in water and then capped toreduce the risk of contamination of the cell sample when removed fromthe large dedicated biosafety cabinet and transported to the integratedbiocontainment cell sorter of the present invention. This reduces therisk of contamination from unwanted foreign material that these cellsamples could be subjected to during transport to the integratedbiocontainment cell sorter. Since the cells are suspended in water,there is a very low risk of accidental exposure to a user. Even when thecap on the sample media is removed, there is low risk of exposure to theuser, since the cells are not in an aerosol form and are suspended inwater. However, the user should wear safety glasses and gloves to reducethe risk of exposure from accidental splashing of the sample into theeyes or mouth. Once the user places the capped sample tubes in thesample input area 107, the tubes can be uncapped in a clean environmentin the sample input area, so that there is a very low likelihood ofcontamination of the sample. Tubes are then placed in the sample inputholders so that a sorting process can commence.

The main cabinet containment system 101, illustrated in FIG. 1 , maytherefore contain dangerous cells in the sample input area 107. Thepartition 114, which may comprise a transparent sliding sash window,allows an operator to easily access the sample input area and the sortchamber 131, through the sort chamber door 132 when the partition 114 isin an upper position in the access opening 115, as shown in FIG. 1 , toinsert and remove samples in the sample input area 107 (FIG. 5 ). Thepartition 114 can be moved to a lower position in the access opening 115to allow direct access by an operator to the nozzle chamber 134, throughnozzle chamber door 136. Fan 122 is sufficiently strong to maintain alow pressure in the work area 104 even though the access opening is onlypartially covered by the partition 114. The partition 114 can simplymove up and down in the access opening 115. Accordingly, the amount ofarea that is closed or blocked off in the access opening 115 by thepartition 114 and the amount of area that is open in the access opening115, and is not closed or blocked off by partition 114, does not changeno matter where the partition 114 is placed in the access opening 115.In other words, the same amount of open area of the access opening 115is present no matter where the partition 114 is located in the accessopening 115. When the partition 114 is located in the up position, asillustrated in FIG. 1 , a certain number of square inches of opening inthe access opening 115 are present. When the partition 114 is moveddownwardly, the amount of open area in the access opening 115 has aconstant size, i.e. the same number of square inches of opening, sincethe partition 114 has a constant size and the access opening 115 has aconstant size. In this manner, the amount of air that is transported byfan 122 can remain the same no matter where the partition 114 is locatedand still maintain a constant first low pressure in the work area 104.In one embodiment, the fan 122 moves about 100 feet of air per minutethrough the user access opening 115 into the work area 104.Functionally, the integrated biocontainment cell sorter system is basedon the velocity of the air that moves through the cabinet. The velocityof the air must be fast enough in order to maintain containment sincethe integrated biocontainment cell sorter is not a sealed system. Inoperation, the air speed of the air that enters the grate 118 ismeasured to ensure proper velocity to maintain containment. The fan 122is designed to operate so that the volume of air that passes through thegrate 118 is sufficient to maintain the containment of hazardousmaterials in the main cabinet containment system 101.

FIG. 2 is a schematic diagram illustrating portions of the aerosolmanagement system (AMS) 149. FIG. 2 specifically illustrates theportions of the cell sorter that are contained within the aerosolmanagement system chambers 140. The nozzle 186, as well as theconnecting tubing, are located in the nozzle chamber 134. The sortplates 176, collection tubes 182, 184 are located in the sort chamber131. Opening 174 allows the droplet stream 155 to flow from the nozzlechamber 134 to the sort chamber 131 through the optics mounting plate150 (FIG. 3 ). FIG. 2 illustrates the primary components that arelocated within the nozzle chamber 134 and the sort chamber 131 thatcomprise the aerosol management system chambers 140. FIG. 2 alsoschematically illustrates the main cabinet containment system 101. Asillustrated in FIG. 2 , the nozzle 186 and the connecting hoses arelocated in the nozzle chamber 134. Sheath fluid container 170 containssheath fluid that is transported to the nozzle 186 via the sheath fluidhose 171. Sheath fluid hose 171 passes through the walls of the maincabinet containment system 101 and the aerosol management systemchambers 140. Containment seals between the sheath fluid hose 171 andthe walls of the main cabinet containment system 101 and the aerosolmanagement system chambers 140 walls are not containment seals thatprovide an airtight seal for the sheath fluid hose 171. Rather, a lessexpensive and easier to install seal can be used since both the maincabinet containment system 101 and the aerosol management systemchambers 140 have low pressures that cause airflow inwardly into themain cabinet containment system 101 and the aerosol management systemchambers 140. The same is true for the sample fluid hose 173.

As also illustrated in FIG. 2 , the aerosol management system chambers140 surround the sort plates, collection tubes 182, 184 and thedeflected droplet streams 156. The nozzle chamber 134 and the sortchamber 131, as well as the opening 174 between these two chambers,comprise the aerosol management system chambers 140. The nozzle 186, theopening 174, the sort plates 176 and the collection tubes 182, 1.84 arethe primary functional components that are contained within the aerosolmanagement system chambers 140. The sample fluid container 172, which isplaced in the sample input area 107 (FIG. 5 ) opens into the maincabinet containment system 101, and not in the aerosol management systemchambers 140. The sheath fluid container 170, the excitation optics 162,the forward scatter detector 180 and the side scatter detector 178 arepreferably all located outside of the main cabinet containment system101. The side scatter light path 168 and the forward scatter light path166 project light through an optical window, as illustrated in FIG. 4 ,to side scatter detector 178 and forward scatter detector 180,respectively. Other electronics and controllers, as well as lasers, arepreferably located outside of the main cabinet containment system 101.For example, as disclosed in U.S. Pat. No. 8,557,587, which isspecifically incorporated herein by reference, for all that it disclosesand teaches, timing and charge circuits, sort logic controllers, opticalfilters, detectors, acquisition electronics, and other electroniccircuits and devices, collectively defined herein as cell sorterelectronics and optical devices, are preferably all located outside ofthe main cabinet and the aerosol management containment area for easyaccess for maintenance and adjustment. In other words, the cell sorterelectronics and optical devices are preferably located in areas that areeasily accessed, and do not require access to contaminated areas withinthe main cabinet containment system 101 or the aerosol management systemchambers 140. Consequently, the excitation optics 162, the side scatterdetector 178 and the forward scatter detector 180 can preferably beeasily accessed without accessing a biocontainment area. Typically,these devices need adjustment, and the accessibility of these devices,without the necessity of entering a dirty or biocontainment area,greatly increases the speed and maintenance of the system.

Of primary importance in easing the maintenance and reducing the size ofa containment system is to have the excitation optics 162 locatedoutside of the main cabinet containment system 101 and the aerosolmanagement system 149. The excitation optics 162 comprises theexcitation lasers or other excitation optics such as LEDs, opticallypumped plasma light generators, arc lamps or other excitation optics.The optics include the various mirrors, beam combiners, lenses, etc.Accordingly, the present invention, in accordance with one embodiment,may simply have the excitation optics 162 located outside of the maincabinet and the other portions, such as the side scatter detector 178,forward scatter detector 180, and other devices located inside of themain cabinet 101 and aerosol management system chambers 140. However,the optical detection devices, such as side scatter detector 178 andforward scatter detector 180, may also be located outside of the maincabinet 101 along with the excitation optics 162, in accordance withanother embodiment of the present invention, and as illustrated in FIG.2 . As a third embodiment of the present invention, the fluidics, suchas the sheath fluid container 170, the various pumps associated with thesheath fluid, may also be located outside of either the aerosolmanagement system chambers 140, or the main cabinet containment system101, either individually or collectively. In other words, variouscombinations of equipment can be located outside of the main cabinet 101and/or the aerosol management system chambers 140 to increaseaccessibility and increase the ease of maintenance of various systemsand also reduce the size of the containment area.

Further, by creating an integrated system, which is specificallyconstructed so that only the nozzle and connecting hoses are in thenozzle chamber 134 and the sort plates 176 and collection tubes 182, 184are in the sort chamber, the volume of the main cabinet containmentsystem 101 is greatly decreased. Many cell sorter systems are simplyplaced in a containment hood, which is very large and bulky. Forexample, containment hoods are typically about nine feet high and can besix or seven feet wide. By only enclosing specific components in thepresent invention, the containment area can be greatly reduced and theoverall size of the integrated biocontainment system 100 can also begreatly reduced. By containing sample fluid container 172 in the inputsample area 107, which is adjacent to the work area 104 of the maincabinet containment system 101, input fluids can be easily inserted andremoved from the input sample area 107 since the sample fluid container172 does not impose as much of a hazard as aerosols that can be createdin the nozzle chamber 134 and the sort chamber 131. In addition, the useof aerosol management system chambers 140 having openings to maincabinet containment system 101 provides additional safety for theoperators since they are not subjected to any of the hazardous particlesthat are contained in the aerosols of the aerosol management systemchambers 140.

FIG. 3 is a schematic cutaway side view of the aerosol management systemchambers 140. As illustrated in FIG. 3 , the nozzle chamber 134 containsthe nozzle 186, which is illustrated in solid lines in the operatingposition. Sheath fluid 170 is provided through the nozzle 186, as wellas the sample fluid 172. The nozzle 186 can also be moved to a cleaningposition, as illustrated in dotted lines. The nozzle chamber door 198can be opened so that the nozzle 186, in the cleaning position, isaccessible through the nozzle chamber door 198. Nozzles, such as nozzle186, can become clogged for various reasons, and a quick and easy accessto the nozzle 186 is provided when the nozzle 186 is in the cleaningposition. Again, when the nozzle chamber door 198 is opened, thenegative pressure in the nozzle chamber 134, compared to the main workarea 104, is equalized so that contaminated air can at that point intime migrate from the nozzle chamber 134 to the main work area 104. Thedroplet stream 155 from the nozzle 186 flows through an opening 152 andthe optics mounting plate 150. The droplet stream 155 passes through aninterrogation point 148 prior to the droplets separating from thestream. Laser beams interrogate the nozzle stream 153 at theinterrogation point 148. Scattered and projected light from theinterrogation point 148 is transmitted through the optical window 200and past the light blocking bar 202 to a side scatter objective 204.Side scatter objective 204 collects the side scatter rays and transmitsthose rays through side scatter pin hole 206. Flexible seals 210, 212and 214 provide a partial seal so that the droplet stream 155 does nottransfer to the non-containment air 208. The droplet stream 155 passesthrough the sort plates 154 and is separated into deflected dropletstreams 156. The AMS intake duct 138 is connected to the AMS HEPA filter141 and the AMS fan 144, as illustrated in FIG. 1 . Air from the sortchamber 131 is drawn through the AMS intake duct 138 to create a lowpressure in both the sort chamber 131 and the nozzle chamber 134. Airfrom the main cabinet work area 104 passes through inlets 188 and 194,Airflow 190 from the main cabinet passes through inlet 188, whileairflow 196 from the main cabinet passes through inlet 194. The sortchamber 131 has a sort chamber door 132 that can be opened to provideoperator access to the sort plates 154 and collection tubes 182, 184(FIG. 2 ) in the sort chamber 131. Again, because the sort chamber 131has lower pressure than the main cabinet work area 104, opening of thesort chamber door 132 equalizes the pressure of the sort chamber and themain cabinet work area 104. To prevent hazardous aerosols from escapingthe sort chamber 131 or nozzle chamber 134, both the sort chamber 131and the nozzle chamber 134 must be evacuated using the AMS fan 144 (FIG.1 ) prior to opening either the nozzle chamber door 198 or the sortchamber door 132. Once the aerosols are purged from the aerosolmanagement chambers 140, the doors 198, 132 can be opened.

FIG. 4 is a front schematic view illustrating the nozzle chamber 134,the sort chamber 131 and various devices of the cell sorter that arelocated outside of the aerosol management system (AMS) 149 and the maincabinet containment system 101. As illustrated in FIG. 4 ,non-containment air 208 surrounds the nozzle chamber wall 213 of thenozzle chamber 134. Laser 216 generates a laser beam along light path164 that passes through an optical window 215. The laser intersects thenozzle stream 153 at interrogation point 148. Light transmitted from theinterrogation point 148 passes through optical window 217 to a lightblocking plate 218 and through the forward scatter objective 220, Theforward scatter objective 220 collects the light and transmits itthrough a pinhole aperture 222 to the light detector 224. The dropletstream 155 passes through an opening 152 in the optics mounting plate150, Flexible seals 228, 230 seal the sort chamber 131 to the opticsmounting plate 150. The droplet stream 155 passes through the sortplates 154 in the sort chamber 131 and is separated by the sort plates154 into deflected droplet streams 156. The sort chamber wall 226separates the sort chamber 131 from a non-containment air region 208.

FIG. 5 is a perspective view of an embodiment of an implementation ofthe integrated biocontainment cell sorter 100. As illustrated in FIG. 5, an exhaust vent 110 exhausts clean air out of the integratedbiocontainment cell sorter 100. Clean air is provided to therecirculation plenum 102 by the recirculation duct 108. The filter andfan plenum 121 contain the filter and fan in a location that is in theback and the bottom of the integrated biocontainment cell sorter 100 sothat other portions of the integrated biocontainment cell sorter 100 areeasily accessible to an operator. Sliding sash window (partition) 114moves with an angular vertical movement to provide an opening to themain cabinet containment system 101. When the sliding sash window 114 isin the upper position, as shown, the sample input area 107 is accessibleby an operator. When the sliding sash window 114 is in a lower position,the nozzle chamber 134 and sample line are accessible. Grate 118 allowsair from outside of the integrated biocontainment cell sorter 100 to bedrawn into the main cabinet containment system 101 so that contaminatedair does not pass out of the main cabinet containment system 101. Bymaintaining a lower pressure in the main cabinet containment system 101,contaminated air does not escape from the integrated biocontainment cellsorter 100.

Accordingly, the integrated biocontainment cell sorter 100 providescontainment only around the portions of the cell sorter that may createcontaminated air. As such, the integrated biocontainment cell sorter 100has containment areas that are small and compact, have smaller fans andsubstantially smaller overall dimensions than cell sorters that areplaced in large hoods or partially integrated biocontainment cellsorters. Partially integrated biocontainment cell sorters encapsulatenumerous components of a cell sorter that do not require containment,and make it much more difficult to provide maintenance to specific areasof the cell sorter that do not require biocontainment. The smallercontainment areas results in smaller fans and requires moving a reducedvolume of air to maintain containment, thereby saving energy.

The foregoing description of the invention has been presented forpurposes of illustration and description. It is not intended to beexhaustive or to limit the invention to the precise form disclosed, andother modifications and variations may be possible in light of the aboveteachings. The embodiment was chosen and described in order to bestexplain the principles of the invention and its practical application tothereby enable others skilled in the art to best utilize the inventionin various embodiments and various modifications as are suited to theparticular use contemplated. It is intended that the appended claims beconstrued to include other alternative embodiments of the inventionexcept insofar as limited by the prior art.

The invention claimed is:
 1. A biocontainment system, comprising: a maincabinet, the main cabinet enclosing a work area therein; a first fantrain, the first fan train comprising a first fan and a first filter,the first fan being operable to draw air from the main cabinet throughthe first filter; an aerosol management system, the aerosol managementsystem comprising a first chamber, the first chamber being locatedwithin the main cabinet and being in fluid communication with the workarea of the main cabinet, the aerosol management system comprising asecond fan train, the second fan train comprising a second fan and asecond filter, the second fan being operable to draw air from the firstchamber through the second filter and to maintain the first chamber at apressure lower than a pressure of the work area.
 2. The biocontainmentsystem of claim 1, wherein the aerosol management system comprises asecond chamber, the second chamber being located within the main cabinetand the second chamber being in fluid communication with the work areaof the main cabinet.
 3. The biocontainment system of claim 2, whereinthe second fan is operable to draw air from the second chamber throughthe second filter and to maintain the second chamber at a pressure lowerthan a pressure of the work area.
 4. The biocontainment system of claim2, comprising a passage between the first chamber and the secondchamber.
 5. The biocontainment system of claim 1, wherein the aerosolmanagement system comprises a recirculation duct, the recirculation ductbeing configured to receive air from the first fan train.
 6. Thebiocontainment system of claim 5, further comprising a recirculationplenum configured to communicate air from the recycle duct to the workarea.
 7. The biocontainment system of claim 5, further comprising anexhaust vent configured to receive air from the first fan train andcommunicate the air to the environment exterior to the main cabinet. 8.The biocontainment system of claim 7, wherein the second fan train is influid communication with the exhaust vent.
 9. The biocontainment systemof claim 1, wherein the main cabinet comprises an opening configured toplace the main cabinet into fluid communication with the environmentexterior to the main.
 10. The biocontainment system of claim 1, furthercomprising an optical system configured to interrogate a sample residingwithin the aerosol management system.
 11. The biocontainment system ofclaim 10, wherein a portion of the optical system is located outside ofthe first chamber.
 12. The biocontainment system of claim 1, furthercomprising a particle sorter, the particle sorter located within theaerosol management system.
 13. A method of containing material in abiocontainment system, comprising: operating a first fan to move airthrough a first flow path, the first flow path being defined by (i) awork area within a main cabinet, (ii) a first filter, and (iii) arecirculation duct in fluid communication with the work area and with afirst exhaust vent; and operating a second fan to move air through anaerosol management system of the biocontainment system, the aerosolmanagement system comprising (i) the second fan, (ii) a first chamber incommunication with the work area, and (iii) a second filter thatreceives air communicated from the first chamber, the aerosol managementsystem being arranged such that the work area is free of air filtered bythe second filter; and maintaining the first chamber at a lower pressurethan the main cabinet.
 14. The method of claim 13, wherein the secondflow path comprises a second chamber, the second chamber beingmaintained at a lower pressure than the main cabinet.
 15. The method ofclaim 13, further comprising sorting a plurality of particles locatedwithin the second flow path.
 16. The method of claim 15, furthercomprising optically exciting the plurality of particles.
 17. The methodof claim 13, wherein the second flow path comprises an exhaust vent. 18.A biocontainment system, comprising: a main cabinet having a work areatherein; a first chamber having a particle processing train locatedtherein, the first chamber being disposed within the main cabinet, thefirst chamber being in fluid communication with the work area; and afiltration train, the filtration train being configured to (i) collectand filter air from the work area, (ii) collect and filter air from thefirst chamber, and (iii) maintain the first chamber at a pressure lowerthan a pressure of the work area.
 19. The biocontainment system of claim18, wherein the particle processing train comprises a particle sorter.20. The biocontainment system of claim 18, wherein the filtration trainis configured to return filtered air to the work area.